Hypoxia stimulates binding of a cytoplasmic protein to a pyrimidine-rich sequence in the 3'-untranslated region of rat tyrosine hydroxylase mRNA
Reduced oxygen tension (hypoxia) induces a 3-fold increase in stability of mRNA for tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis, in the pheochromocytoma (PC12) clonal cell line. To investigate the possibility that RNA-protein interactions are involved in mediating...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 1994-04, Vol.269 (13), p.9940-9945 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 9945 |
|---|---|
| container_issue | 13 |
| container_start_page | 9940 |
| container_title | The Journal of biological chemistry |
| container_volume | 269 |
| creator | CZYZYK-KRZESKA, M. F DOMINSKI, Z KOLE, R MILLHORN, D. E |
| description | Reduced oxygen tension (hypoxia) induces a 3-fold increase in stability of mRNA for tyrosine hydroxylase (TH), the rate-limiting
enzyme in catecholamine synthesis, in the pheochromocytoma (PC12) clonal cell line. To investigate the possibility that RNA-protein
interactions are involved in mediating this increase in stability, RNA gel shift assays were performed using different fragments
of labeled TH mRNA and the S-100 fraction of PC12 cytoplasmic protein extracts. We identified a sequence within the 3'-untranslated
region of TH mRNA that binds cytoplasmic protein. RNase T1 mapping revealed that the protein was bound to a 28 nucleotide
long sequence that is located between bases 1551-1579 of TH mRNA. Moreover, protein binding to this fragment was prevented
with an antisense oligonucleotide directed against bases 1551-1579 and subsequent RNase H digestion. This fragment of the
3'-untranslated region of TH mRNA is rich in pyrimidine nucleotides, and the binding of cytoplasmic protein to this fragment
was reduced by competition with other polypyrimidine sequences including poly(C) but not poly(U) polymers. The binding of
the protein to TH mRNA was increased when cytoplasmic proteins were extracted from PC12 cells exposed to hypoxia (5% O2) for
24 h. Electrophoresis of the UV cross-linked RNA-protein complex on SDS-polyacrylamide gel electrophoresis revealed a complex
of 74 kDa. The potential role of this protein-TH mRNA interaction in regulation of TH mRNA stability during hypoxia is discussed. |
| doi_str_mv | 10.1016/S0021-9258(17)36973-9 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_76419814</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>76419814</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3549-51647979d3192ed8c3d6a339347d604a1eee9a4ac2b6f236742e23abe0eaafdd3</originalsourceid><addsrcrecordid>eNqFkc1u1TAQhS0EKreFR6hkCURhEbBjx46XVQUUqQKJH4md5diTG6MkDnYimrfgkXF6r-4Wb7w438yZmYPQJSVvKaHi3TdCSlqosqpfU_mGCSVZoR6hHSU1K1hFfz5GuxPyFJ2n9IvkxxU9Q2dSkbqs1Q79vV2ncO8NTrMflt7MkHDjR-fHPQ4tNtiuc5h6kwZv8RTDDH7Ec8jCtEY_-AxCEb3tcILfC4wW8AZ0gNlVsYxzNGPaujocYe_DuDWNZsbzGkPKtbhbXQz3a3YAPHz9fP0MPWlNn-D58b9APz68_35zW9x9-fjp5vqusKziqqio4FJJ5RhVJbjaMicMY4px6QThhgKAMtzYshFtyYTkJZTMNEDAmNY5doFeHfrmpfLgadaDTxb63owQlqSl4FTVlP8XpKIWkhKSweoA2rxaitDqKV_IxFVTorfI9ENkestDU6kfItMq110eDZZmAHeqOmaU9ZdH3SRr-jaf1Pp0wjilSsptzhcHrPP77o-PoBsfbAeDLoXSNDspTtg_YW2suQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16867100</pqid></control><display><type>article</type><title>Hypoxia stimulates binding of a cytoplasmic protein to a pyrimidine-rich sequence in the 3'-untranslated region of rat tyrosine hydroxylase mRNA</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>CZYZYK-KRZESKA, M. F ; DOMINSKI, Z ; KOLE, R ; MILLHORN, D. E</creator><creatorcontrib>CZYZYK-KRZESKA, M. F ; DOMINSKI, Z ; KOLE, R ; MILLHORN, D. E</creatorcontrib><description>Reduced oxygen tension (hypoxia) induces a 3-fold increase in stability of mRNA for tyrosine hydroxylase (TH), the rate-limiting
enzyme in catecholamine synthesis, in the pheochromocytoma (PC12) clonal cell line. To investigate the possibility that RNA-protein
interactions are involved in mediating this increase in stability, RNA gel shift assays were performed using different fragments
of labeled TH mRNA and the S-100 fraction of PC12 cytoplasmic protein extracts. We identified a sequence within the 3'-untranslated
region of TH mRNA that binds cytoplasmic protein. RNase T1 mapping revealed that the protein was bound to a 28 nucleotide
long sequence that is located between bases 1551-1579 of TH mRNA. Moreover, protein binding to this fragment was prevented
with an antisense oligonucleotide directed against bases 1551-1579 and subsequent RNase H digestion. This fragment of the
3'-untranslated region of TH mRNA is rich in pyrimidine nucleotides, and the binding of cytoplasmic protein to this fragment
was reduced by competition with other polypyrimidine sequences including poly(C) but not poly(U) polymers. The binding of
the protein to TH mRNA was increased when cytoplasmic proteins were extracted from PC12 cells exposed to hypoxia (5% O2) for
24 h. Electrophoresis of the UV cross-linked RNA-protein complex on SDS-polyacrylamide gel electrophoresis revealed a complex
of 74 kDa. The potential role of this protein-TH mRNA interaction in regulation of TH mRNA stability during hypoxia is discussed.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(17)36973-9</identifier><identifier>PMID: 7908289</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: American Society for Biochemistry and Molecular Biology</publisher><subject>Analytical, structural and metabolic biochemistry ; Animals ; Base Sequence ; Binding Sites ; Biological and medical sciences ; Cell Hypoxia - physiology ; Cells, Cultured ; Cytoplasm - metabolism ; DNA, Complementary - metabolism ; Enzymes and enzyme inhibitors ; Fundamental and applied biological sciences. Psychology ; Molecular Sequence Data ; Oligonucleotides, Antisense ; Oxidoreductases ; PC12 Cells ; Protein Binding ; Protein Biosynthesis ; Rats ; Restriction Mapping ; RNA, Messenger - isolation & purification ; RNA, Messenger - metabolism ; RNA-Binding Proteins - isolation & purification ; RNA-Binding Proteins - metabolism ; Transcription, Genetic ; Tyrosine 3-Monooxygenase - biosynthesis</subject><ispartof>The Journal of biological chemistry, 1994-04, Vol.269 (13), p.9940-9945</ispartof><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3549-51647979d3192ed8c3d6a339347d604a1eee9a4ac2b6f236742e23abe0eaafdd3</citedby><cites>FETCH-LOGICAL-c3549-51647979d3192ed8c3d6a339347d604a1eee9a4ac2b6f236742e23abe0eaafdd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27922,27923</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4119774$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7908289$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>CZYZYK-KRZESKA, M. F</creatorcontrib><creatorcontrib>DOMINSKI, Z</creatorcontrib><creatorcontrib>KOLE, R</creatorcontrib><creatorcontrib>MILLHORN, D. E</creatorcontrib><title>Hypoxia stimulates binding of a cytoplasmic protein to a pyrimidine-rich sequence in the 3'-untranslated region of rat tyrosine hydroxylase mRNA</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Reduced oxygen tension (hypoxia) induces a 3-fold increase in stability of mRNA for tyrosine hydroxylase (TH), the rate-limiting
enzyme in catecholamine synthesis, in the pheochromocytoma (PC12) clonal cell line. To investigate the possibility that RNA-protein
interactions are involved in mediating this increase in stability, RNA gel shift assays were performed using different fragments
of labeled TH mRNA and the S-100 fraction of PC12 cytoplasmic protein extracts. We identified a sequence within the 3'-untranslated
region of TH mRNA that binds cytoplasmic protein. RNase T1 mapping revealed that the protein was bound to a 28 nucleotide
long sequence that is located between bases 1551-1579 of TH mRNA. Moreover, protein binding to this fragment was prevented
with an antisense oligonucleotide directed against bases 1551-1579 and subsequent RNase H digestion. This fragment of the
3'-untranslated region of TH mRNA is rich in pyrimidine nucleotides, and the binding of cytoplasmic protein to this fragment
was reduced by competition with other polypyrimidine sequences including poly(C) but not poly(U) polymers. The binding of
the protein to TH mRNA was increased when cytoplasmic proteins were extracted from PC12 cells exposed to hypoxia (5% O2) for
24 h. Electrophoresis of the UV cross-linked RNA-protein complex on SDS-polyacrylamide gel electrophoresis revealed a complex
of 74 kDa. The potential role of this protein-TH mRNA interaction in regulation of TH mRNA stability during hypoxia is discussed.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Binding Sites</subject><subject>Biological and medical sciences</subject><subject>Cell Hypoxia - physiology</subject><subject>Cells, Cultured</subject><subject>Cytoplasm - metabolism</subject><subject>DNA, Complementary - metabolism</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Molecular Sequence Data</subject><subject>Oligonucleotides, Antisense</subject><subject>Oxidoreductases</subject><subject>PC12 Cells</subject><subject>Protein Binding</subject><subject>Protein Biosynthesis</subject><subject>Rats</subject><subject>Restriction Mapping</subject><subject>RNA, Messenger - isolation & purification</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA-Binding Proteins - isolation & purification</subject><subject>RNA-Binding Proteins - metabolism</subject><subject>Transcription, Genetic</subject><subject>Tyrosine 3-Monooxygenase - biosynthesis</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1u1TAQhS0EKreFR6hkCURhEbBjx46XVQUUqQKJH4md5diTG6MkDnYimrfgkXF6r-4Wb7w438yZmYPQJSVvKaHi3TdCSlqosqpfU_mGCSVZoR6hHSU1K1hFfz5GuxPyFJ2n9IvkxxU9Q2dSkbqs1Q79vV2ncO8NTrMflt7MkHDjR-fHPQ4tNtiuc5h6kwZv8RTDDH7Ec8jCtEY_-AxCEb3tcILfC4wW8AZ0gNlVsYxzNGPaujocYe_DuDWNZsbzGkPKtbhbXQz3a3YAPHz9fP0MPWlNn-D58b9APz68_35zW9x9-fjp5vqusKziqqio4FJJ5RhVJbjaMicMY4px6QThhgKAMtzYshFtyYTkJZTMNEDAmNY5doFeHfrmpfLgadaDTxb63owQlqSl4FTVlP8XpKIWkhKSweoA2rxaitDqKV_IxFVTorfI9ENkestDU6kfItMq110eDZZmAHeqOmaU9ZdH3SRr-jaf1Pp0wjilSsptzhcHrPP77o-PoBsfbAeDLoXSNDspTtg_YW2suQ</recordid><startdate>19940401</startdate><enddate>19940401</enddate><creator>CZYZYK-KRZESKA, M. F</creator><creator>DOMINSKI, Z</creator><creator>KOLE, R</creator><creator>MILLHORN, D. E</creator><general>American Society for Biochemistry and Molecular Biology</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>19940401</creationdate><title>Hypoxia stimulates binding of a cytoplasmic protein to a pyrimidine-rich sequence in the 3'-untranslated region of rat tyrosine hydroxylase mRNA</title><author>CZYZYK-KRZESKA, M. F ; DOMINSKI, Z ; KOLE, R ; MILLHORN, D. E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3549-51647979d3192ed8c3d6a339347d604a1eee9a4ac2b6f236742e23abe0eaafdd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Binding Sites</topic><topic>Biological and medical sciences</topic><topic>Cell Hypoxia - physiology</topic><topic>Cells, Cultured</topic><topic>Cytoplasm - metabolism</topic><topic>DNA, Complementary - metabolism</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Molecular Sequence Data</topic><topic>Oligonucleotides, Antisense</topic><topic>Oxidoreductases</topic><topic>PC12 Cells</topic><topic>Protein Binding</topic><topic>Protein Biosynthesis</topic><topic>Rats</topic><topic>Restriction Mapping</topic><topic>RNA, Messenger - isolation & purification</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA-Binding Proteins - isolation & purification</topic><topic>RNA-Binding Proteins - metabolism</topic><topic>Transcription, Genetic</topic><topic>Tyrosine 3-Monooxygenase - biosynthesis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>CZYZYK-KRZESKA, M. F</creatorcontrib><creatorcontrib>DOMINSKI, Z</creatorcontrib><creatorcontrib>KOLE, R</creatorcontrib><creatorcontrib>MILLHORN, D. E</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>CZYZYK-KRZESKA, M. F</au><au>DOMINSKI, Z</au><au>KOLE, R</au><au>MILLHORN, D. E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Hypoxia stimulates binding of a cytoplasmic protein to a pyrimidine-rich sequence in the 3'-untranslated region of rat tyrosine hydroxylase mRNA</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1994-04-01</date><risdate>1994</risdate><volume>269</volume><issue>13</issue><spage>9940</spage><epage>9945</epage><pages>9940-9945</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>Reduced oxygen tension (hypoxia) induces a 3-fold increase in stability of mRNA for tyrosine hydroxylase (TH), the rate-limiting
enzyme in catecholamine synthesis, in the pheochromocytoma (PC12) clonal cell line. To investigate the possibility that RNA-protein
interactions are involved in mediating this increase in stability, RNA gel shift assays were performed using different fragments
of labeled TH mRNA and the S-100 fraction of PC12 cytoplasmic protein extracts. We identified a sequence within the 3'-untranslated
region of TH mRNA that binds cytoplasmic protein. RNase T1 mapping revealed that the protein was bound to a 28 nucleotide
long sequence that is located between bases 1551-1579 of TH mRNA. Moreover, protein binding to this fragment was prevented
with an antisense oligonucleotide directed against bases 1551-1579 and subsequent RNase H digestion. This fragment of the
3'-untranslated region of TH mRNA is rich in pyrimidine nucleotides, and the binding of cytoplasmic protein to this fragment
was reduced by competition with other polypyrimidine sequences including poly(C) but not poly(U) polymers. The binding of
the protein to TH mRNA was increased when cytoplasmic proteins were extracted from PC12 cells exposed to hypoxia (5% O2) for
24 h. Electrophoresis of the UV cross-linked RNA-protein complex on SDS-polyacrylamide gel electrophoresis revealed a complex
of 74 kDa. The potential role of this protein-TH mRNA interaction in regulation of TH mRNA stability during hypoxia is discussed.</abstract><cop>Bethesda, MD</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>7908289</pmid><doi>10.1016/S0021-9258(17)36973-9</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 1994-04, Vol.269 (13), p.9940-9945 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_76419814 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Analytical, structural and metabolic biochemistry Animals Base Sequence Binding Sites Biological and medical sciences Cell Hypoxia - physiology Cells, Cultured Cytoplasm - metabolism DNA, Complementary - metabolism Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology Molecular Sequence Data Oligonucleotides, Antisense Oxidoreductases PC12 Cells Protein Binding Protein Biosynthesis Rats Restriction Mapping RNA, Messenger - isolation & purification RNA, Messenger - metabolism RNA-Binding Proteins - isolation & purification RNA-Binding Proteins - metabolism Transcription, Genetic Tyrosine 3-Monooxygenase - biosynthesis |
| title | Hypoxia stimulates binding of a cytoplasmic protein to a pyrimidine-rich sequence in the 3'-untranslated region of rat tyrosine hydroxylase mRNA |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-09T14%3A55%3A11IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Hypoxia%20stimulates%20binding%20of%20a%20cytoplasmic%20protein%20to%20a%20pyrimidine-rich%20sequence%20in%20the%203'-untranslated%20region%20of%20rat%20tyrosine%20hydroxylase%20mRNA&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=CZYZYK-KRZESKA,%20M.%20F&rft.date=1994-04-01&rft.volume=269&rft.issue=13&rft.spage=9940&rft.epage=9945&rft.pages=9940-9945&rft.issn=0021-9258&rft.eissn=1083-351X&rft.coden=JBCHA3&rft_id=info:doi/10.1016/S0021-9258(17)36973-9&rft_dat=%3Cproquest_cross%3E76419814%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=16867100&rft_id=info:pmid/7908289&rfr_iscdi=true |